Hyperparasitism in bat flies (Diptera: Streblidae): new records and interaction networks in the Neotropics.

Hyperparasitism is defined as the interaction where one parasite is infected by another parasite. In bat flies (Streblidae and Nycteribiidae), both hyperparasites and microparasites (bacteria, viruses, fungi, and arthropods such as mites) have been documented. Fungi belonging to the order Laboulbeniales are microscopic parasites of a wide diversity of arthropod hosts. Three genera exclusively target bat flies: Arthrorhynchus, which parasitizes species within Nycteribiidae in the Eastern Hemisphere, while genus Gloeandromyces and Nycteromyces parasitize Streblidae in the Western Hemisphere. Among the hyperparasitic arthropods, mites of family Neothrombidiidae, particularly the monospecific genus Monunguis, are known to parasitize bat flies. Here we present the first records of the hyperparasites Monunguis streblida and Gloeandromyces pageanus f. polymorphus parasitizing Streblidae bat flies in Colombia and a summary of these hyperparasitic interactions in the Neotropics. We detected fungi and mites parasitizing bat flies that were collected in the Magdalena River Basin, Colombia, in field expeditions in 2018, 2022, and 2023. We identified 17 bat flies and two species of hyperparasites, specifically M. streblida and the fungi Gloeandromyces. Our search for reports of these interactions in the Neotropics revealed that seven species of Trichobius (Streblidae) are parasitized by M. streblida, whereas Paratrichobius longicrus (Streblidae) is parasitized by Gloeandromyces pageanus f. polymorphus. These interactions have been reported in 11 countries, but our records are the first of M. streblida and Laboulbeniales fungi parasitizing bat flies in Colombia. So far, a total of 14 species of fungi and one species of mite have been associated with 19 species of bat flies, which in turn, are linked to 15 species of Neotropical bats.

Sarcoptic mange in wild quichua porcupines (Coendou quichua Thomas, 1899) in Colombia.

The Quichua porcupine (Coendou quichua) is a neotropical rodent with uncertain taxonomic and conservation status. Two Quichua porcupines with severe hyperkeratosis and alopecia were found in the Magdalena River Basin of Colombia. Sarcoptes scabiei, the mite causing mange, a disease carried mainly by domestic animals, was confirmed via parasitological and molecular methods. This is the first report of mange in neotropical porcupines to date. The population-level impact of mange in Coendou spp., related mammals and predators in Colombia might represent a threat and needs further investigation.

Comparative analysis of germ cells and DNA of the genus Amblyomma: adding new data on Amblyomma maculatum and Amblyomma ovale species (Acari: Ixodidae).

Among tick species, members of the subfamily Amblyomminae have received special attention, since they serve as vectors for pathogens such as Rickettsia spp. and display cryptic species complexes that make their taxonomical classification challenging. Amblyomma ovale, Amblyomma maculatum, and other species of the genus Amblyomma have shown a long history of taxonomic controversies. Spermiotaxonomy has proved to be a valuable tool in the solution of systematic conflicts in Metazoa that can aid molecular and external morphological analyses in ticks and, overall, provide more robust analyses and results. With this in mind, this study included histological analyses of the reproductive system of the species A. ovale and A. maculatum, as well as the description of morphohistological characters of the male reproductive system of ticks of the genus Amblyomma, in order to evaluate these characters within the current clustering proposals. In addition, 16S rDNA and COI (mitochondrial) molecular markers were used to study the genetic relationships of the species. The results show that the tick male reproductive system and its germ cells contain useful candidate characters for taxonomical analyses of Ixodida.

COMPARACIÓN DE MÉTODOS CONVENCIONALES Y MOLECULARES PARA LA DETECCIÓN DE Giardia lamblia EN HECES HUMANAS / COMPARISON OF CONVENTIONAL AND MOLECULAR METHODS FOR DETECTION OF Giardia lamblia IN HUMAN FEACES

Los protozoarios del género Giardia representan algunos de los parásitos humanos más comunes en el mundo y están entre los principales causantes de infecciones gastrointestinales y enfermedades diarreicas en humanos. La detección del parásito se fundamenta generalmente en los métodos por concentración y microscopía convencional, pero estas técnicas presentan limitaciones por su baja sensibilidad e inespecificidad en el diagnóstico. En procura de mejorar los métodos de diagnóstico, las técnicas moleculares se perfilan como una alternativa promisoria. En este estudio se analizaron 88 muestras de heces provenientes de pacientes de una empresa prestadora de servicios de salud (ASSBASALUD) de la ciudad de Manizales (Caldas). Para la detección de Giardia lamblia en heces se compararon tres métodos diferentes por medio del porcentaje de positividad: los métodos convencionales de concentración de la muestra y observación microscópica, análisis por inmunoensayo (ELISA indirecto) y finalmente la amplificación de dos secuencias génicas nucleares por PCR. Se obtuvieron tres muestras positivas por concentración y microscopía convencional, dos por inmunoensayo y 26 por técnicas moleculares. El estudio sugiere que las pruebas diagnósticas rutinarias basadas en microscopía convencional e inmunoensayo, tienen más bajo porcentaje de detección de este parásito y que esta deficiencia puede ser compensada por medio de la implementación de métodos de diagnóstico molecular basados en PCR, como una estrategia complementaria de apoyo en el diagnóstico de este protozoo.

The protozoa of the genus Giardia represent one of the most common human parasites in the world and are among the main causes of gastrointestinal infections and diarrheal diseases in humans. Parasite detection is generally based on concentration and conventional microscopy methods, but these techniques have limitations due to their low sensitivity and specificity in diagnosis. In an attempt to improve the diagnostic methods, molecular techniques are emerging as a promising alternative. In this study 88 stool samples from patients of a company that provides health services (ASSBASALUD) in the city of Manizales (Caldas) were analyzed. In order to detect Giardia lamblia in stool, three different methods were compared using the positivity percent: conventional methods for concentration of the sample and microscopic observation, analysis by immunoassay (indirect ELISA) and finally the amplification of two nuclear gene sequences by PCR. Three positive samples were obtained by concentration and conventional microscopy, two by immunoassay and 26 by molecular techniques. The study suggests that routine diagnostic tests based on conventional microscopy and immunoassay have lower detection rate of this parasite and that this deficiency can be compensated by means of the implementation of molecular diagnostic methods based on PCR, as a complementary strategy to support the diagnosis of this protozoan.

EVALUACIÓN DE MÉTODOS MOLECULARES Y MICROSCÓPICOS PARA LA DETECCIÓN DE Cryptosporidium spp. (APICOMPLEXA - CRYPTOSPORIDIIDAE) / EVALUATION OF MOLECULAR AND MICROSCOPIC METHODS FOR DETECTION OF Cryptosporidium spp. (APICOMPLEXA -CRYPTOSPORIDIIDAE)

Introducción. La criptosporidiosis es una enfermedad emergente causada por protozoarios del género Cryptosporidium, afecta un amplio rango de vertebrados incluyendo al hombre, su prevalencia oscila entre el 4-6% en centro y sur América y puede llegar a causar la muerte en pacientes inmunosuprimidos, por lo que es considerada un problema de salud pública en todo el mundo. Se hace necesario implementar y evaluar estrategias de detección y tipificación de las distintas especies de Cryptosporidium, para adoptar medidas de control y seguimiento. Objetivo. Realizar una comparación de métodos microscópicos y moleculares para la detección y tipificación de las especies de Cryptosporidium, con el fin de utilizar aquel de mayor sensibilidad en la detección del parásito en muestras de agua. Materiales y métodos. La detección y tipificación de Cryptosporidium spp., en muestras fecales y de agua, usando inicialmente un método de concentración tanto para las heces como para el agua (formol-éter y el método de floculación inorgánica con carbonato de calcio); la identificación del parásito se realizó por la tinción de Ziehl-Neelsen y la amplificación por Reacción en Cadena de la Polimerasa (PCR) de regiones del ADN ribosomal, de genes que codifican para la proteína Hsp70 y del gen que codifica para la proteína de la pared del ooquiste de Cryptosporidium (COWP). La tipificación se realizó por medio de digestión con las enzimas de restricción SspI, VspI y RsaI. Resultados. La tinción de Ziehl-Neelsen, comprobó la presencia de Cryptosporidium spp., en 10 de las 168 muestras analizadas (humanos, terneros, perros y conejos), la tipificación por PCR, confirmaron 15 muestras positivas para C. parvum y una para C. hominis. Conclusiones. Se demuestra la sensibilidad de la detección de Cryptosporidium, por PCR y su utilidad en el diagnóstico, al registrar la presencia de dos especies del parásito circulando en muestras del municipio de Manizales.

Introduction. Cryptosporidiosis is an emerging disease caused by protozoa of the Cryptosporidium genus,which affects a wide range of vertebrates, including humansIts prevalence ranges from 4%-6% in Central and South America and can even cause death in immunosuppressed patients reason why it is considered a public health problem worldwide. It is necessary to implement and evaluate strategies for detection and typification of different Cryptosporidium species,to adopt control measures and monitoring. Objective. To perform a comparison between microscopic and molecular methods for the detection and typification of Cryptosporidium species with the purpose of using the most sensitive method in the detection of Cryptosporidium oocysts in water samples. Materials and methods. Detection and typification of Cryptosporidium spp, in feces and water samples were done initially using a concentration method for both feces and water (Formol-ether and inorganic flocculation method with Calcium carbonate); the parasite identification was carried out using the Ziehl-Nelseen staining and the Polymerase Chain Reaction PCR amplification of ribosomal ADN regions, of gens codified for Hsp70 protein and the gen that codifies for the Cryptosporidium oocyst wall protein (COWP). The typification was carried out by digestion with restriction enzymes SspI, RsaI and VspI. Results. The Ziehl-Neelsen staining, confirmed the presence of Cryptosporidium spp., in 10 of the 168 samples tested (humans, calves, dogs and rabbits), the PCR typification, confirmed 15 positive samples for C. parvum and one for C. hominis. Conclusion. The sensitivity of detection of Cryptosporidium by PCR and its utility in the diagnosis is shown, registering the presence of two species of the parasite circulating in samples taken in the municipality of Manizales.

CARACTERIZACIÓN MOLECULAR DE CLONES DE Theobroma cacao L, POR MEDIO DE MARCADORES MOLECULARES MICROSATÉLITES / Theobroma cacao L. CLONES MOLECULAR CHARACTERIZATION USING MOLECULAR MICROSATELLITE MARKERS

Theobroma cacao L. es la única especie del género Theobroma que se explota comercialmente en grandes extensiones, registrando en la actualidad una amplia distribución mundial, a través de programas de desarrollo directamente influenciados por factores vinculados al mercado y por los intereses de productores, comerciantes, industriales y consumidores. En este estudio se evaluaron 12 clones de cacao por medio de marcadores moleculares utilizando 10 secuencias microsatélites (SSRs), como un estudio piloto antes de un ensayo a mayor escala. Los datos se procesaron mediante el programa Power Marker Versión 3.25. Se estimaron las frecuencias alélicas y luego se generó una matriz de distancias genéticas basada en el coeficiente de Nei. Utilizando el algoritmo de agrupamiento UPGMA se generó el dendrograma correspondiente. El análisis de diversidad mostró un índice de diversidad genética total de 0,6944 considerado intermedio para los materiales evaluados. La estimación de la heterocigosidad promedio fue de 0,58579 y de promedio del contenido de información polimórfica de 0,6523. En este estudio los marcadores mTcCIR6, mTcCIR25, mTcCIR26 y mTcCIR12 son los más informativos y polimórficos. Se recomienda relacionar los resultados de diversidad genética con caracteres morfo-agronómicos y de patogenicidad en los distintos clones de cacao a fin de consolidar estrategias eficaces de mejoramiento y control de enfermedades.

Theobroma cacao L. is the only Theobroma species that is commercially exploited in enormous extensions, registering presently wide distribution worldwide through development programs directly influenced by factors linked to the market and the producers, traders, industrials, and consumers’ interests. In this study 12 cacao clones were evaluated by means of molecular markers using 10 microsatellite sequences (SSRs) as a pilot study before a test at a greater scale. The data were processed using the Power Maker Version 3.25 program. The allelic frequencies were stimated and a genetic distance matrix was developed based on the Nei coefficient. Using the UPGMA grouping algorithm the corresponding dendrogram was generated. The diversity analysis showed a total genetic diversity index of 0.6944 considered intermediate for the materials evaluated. The average heterozygocity estimate was 0.58579 and the average polymorphic information content was 0.6523. In this study markers mTcCIR6, mTcCIR25, mTcCIR26 and mTcCIR12 are the most informative and polymorphic ones. It is recommended to relate the genetic diversity results with morpho-agronomic and pathogenicity characters in the different cacao clones in order to consolidate efficient strategies for disease improvement and control.